Antiviral and comma antibacterial pharmaceutical compositions of cantharidic anhydride and method of preparation thereof

ABSTRACT

The present invention relates to an antiviral and antibacterial pharmaceutical composition comprising cantharidic anhydride-essential oil preparation as effective ingredient and method for preparing thereof. The cantharidic anhydride-essential oil preparation is prepared by dissolving cantharidic anhydride at appropriate temperatures. The clinical trials demonstrate that the cantharidic anhydride-essential oil preparation is effective in the treatment of tuberculosis and various viruses infected diseases. Furthermore, a series of local formulations for external use for treatment and prevention of virus infections can be prepared from the said cantharidic anhydride-essential oil preparations.

CROSS REFERENCE TO RELATED APPLICATION(S)

This application is the National Stage of International Application No.PCT/CN02/00839 filed Nov. 22, 2002, which in turn claims the benefit ofChinese Application No. 01140066.8 filed Nov. 23, 2001.

FIELD OF THE INVENTION

The present invention provides antiviral and antibacterialpharmaceutical compositions of cantharidic anhydride and method forpreparing thereof. More specifically, the invention provides antiviraland antibacterial pharmaceutical composition comprising cantharidicanhydride-essential oils preparation obtained by dissolving acantharidic anhydride in essential oils or single constituent ofessential oils as the effective ingredient and pharmaceuticallyacceptable vehicle or excipient. The clinical trials demonstrate thatthe pharmaceutical composition of present invention has excellentantibacterial and antiviral effect, particularly is effective in thetreatment of the diseases infected by tubercule bacillus,staphylococcus, hepatitis virus, human immunodeficiency virus, influenzavirus and rotavirus.

BACKGROUND OF THE INVENTION

Cantharidic anhydride (cantharidin, hydroxyl cantharidin) is anextremely poisonous substance extracted from the body of cantharis,cantharid and the like used in Chinese traditional medicine. Cantharisused as a drug was recorded to have anticancer effect in Chinesetraditional medicine long ago. Classical cantharis plaster abroad is ablister formed by combining cantharis powder with bee wax, rosin, tallowetc. In the said plaster, the content of cantharis powder was 25%.According to solubility calculation and test observation, most of thecantharidic anhydride was not dissolved in the course of plasterpreparation. Therefore this classical skin-irritating agent was only amanufactured product of cantharis powder. In recent years, especiallyafter the discovery of that cantharidic anhydride can serve asanticancer drugs, studies on the pharmacology and clinical applicationof cantharidic anhydride grows rapidly. At present, cantharidicanhydride is obtained by the extraction of cantharis powder by organicsolvents such as acetone, chloroform, ether and the like. In order toimprove the yield of cantharidic anhydride from the conversion ofmagnesium cantharidate, people usually treated cantharis powder withconcentrated hydrochloric acid before the extraction of cantharidicanhydride with organic solvents. The obtained cantharidic anhydride orhydroxyl cantharidic acid were then purified and made intopharmaceutical composition, for example, cantharidin tablet (ShanghaiQ/WS-1-566-80) and cantharidin injection (Shanghai Q/WS-1-567-80) havebeen granted.

Recently it was found that cantharidic anhydride is an antiviralantibiotics. However, in human body and animal body, the said antibioticeffect of cantharidic anhydride is not obvious for non-fat-solublespecies but is quite well for fat-soluble cantharidic anhydride (Journalof Nature, 1980, Vol. 3, 458 in Chinese), especially in the case oftreating viral hepatitis. Preparations made from fat-soluble cantharidicanhydride are specific for such diseases. The present inventor hasalready disclosed the preparation of cantharidic anhydride in his earlyChinese Patent Application (ZL 90106365.7). The preparation for thefat-soluble cantharidic anhydride pharmaceutical composition mainlycomprises the following steps:

-   -   1. Firstly cantharis powder was soaked in organic solvents such        as acetone, dichloromethane, trichloromethane and ether to        extract the cantharidic oil and cantharidic anhydride in the        cantharis powder into the organic phase.    -   2. After the cantharis powder was extracted with organic        solvent, the residue was dried and then was soaked in solution        of concentrate hydrochloric acid and acetone. Cantharidic acid        is converted into cantharidic anhydride by concentrated        hydrochloric acid and cantharidic anhydride is dissolved into        organic phase by using acetone.    -   3. After the cantharidic anhydride and cantharidic oil in the        above-mentioned organic phase were separated from the organic        phase, petroleum ether was added to extract the cantharidic oil        and thus the separation of cantharidic anhydride from        cantharidic oil was achieved. The cantharidic anhydride obtained        was combined with that separated in step 2.    -   4. A definite amount of cantharidic oil was added to refined fat        and then a amount of cantharidic anhydride was added and the        mixture was stirred and heated at 80-120° C. and finally        intermediate product of fat soluble cantharidic anhydride was        obtained.

The above-mentioned refined fat can be vegetable oils such as soy beanoil, corn oil vegetable seed oil, sun flower seed oil, sesame oil, cocoabutter and etc, or wool fat, tallow and the like.

The drugs containing fat solvent of cantharidic anhydride in theabove-mentioned patent having good application value. However, the kindof formulation is limited by the nature of fat solvent and drug. Forexample, the preparatory technology and clinical operation of theinjection made of fat solvent are not satisfied and convenient.

On the other hand, one third of the effective components of cantharidicanhydride in the body of cantharis exists in the form of salt ofcantharidic acid. Therefore, in traditional Chinese medicine, the effectof medicine also comprises that from magnesium cantharidate whencantharis body was used as oral medication. Since salt of cantharidicacid is easily soluble in water, it would have advantage in preparatorytechnology and clinical operation. The method of extraction of magnesiumcantharidate or salt of cantharidic acid from the body of cantharis andconvertion them into purified product are quite complex and have a lowyield. It is much more simple, economic, convenient to purifycantharidic anhydride first and then prepare pure salt of cantharidicacid through semi-synthesis. The existence of magnesium cantharidate andsalt of cantharidic acid in natural cantharis body indicates that theyexhibit biological antibiotic effect in the living cantharis body.Compared with cantharidic anhydride, they have the advantage of highersolubility in the body fluid of cantharis. Therefore salt of cantharidicacid has its own character to be an antibacterial and antiviral drug.

In addition, antiviral preparations containing salt of cantharidic acidare convenient to use, have antibiotic effect to many kinds of importantbacteria and viruses and have applicability to wide range of medicalpractices. Several kinds of pharmaceutical preparations made of salt ofcantharidic acid have similar antiviral effect as fatsolvent-cantharidic anhydride. Pharmaceutical preparations made of saltof cantharidic acid can be used to certain fields of medical treatmentand disease prevention that fat solvent-cantharidic anhydride can not beused.

The above-mentioned technology was disclosed in detail in another patent(ZL931101573) of the present inventor. However the preparatorytechnology for pharmaceutical preparations of salt of cantharidic acidis more complex and thus the manufacturing cost is higher.

Besides, as mentioned above, although cantharidic anhydride hasantiviral effect, there is not any report regarding antibacterial effectfor bacteria such as tubercule bacillus.

CONTENT OF THE INVENTION

The objective of the present invention is to provide an antiviral andantibacterial pharmaceutical composition. The pharmaceutical compositionhas excellent antibacterial and antiviral effect on pathogenicmicroorganisms such as tubercule bacillus, staphylococcus, hepatitisvirus, human immunodeficiency virus (HIV), influenza virus androtavirus. In addition, the present invention also provides an antiviraland antibacterial pharmaceutical composition for external local use.Preparations containing the pharmaceutical composition for local usescan widely be used for sterilization, sanitation and health care.

Another objective of the present invention is to provide method ofpreparation for the above-mentioned pharmaceutical composition.

To achieve above objective, the present invention provides an antiviraland antibacterial pharmaceutical composition comprising cantharidicanhydride as the effective ingredient and pharmaceutically acceptablevehicle or excipient, wherein cantharidic anhydride is dissolved inessential oils or single constituent of essential oils to formcantharidic anhydride-essential oils preparation, and the saidcantharidic anhydride is in a range of 0.05%-0.7% by weight based ontotal weight of cantharidic anhydride-essential oils preparation.

Besides, the antiviral and antibacterial pharmaceutical composition ofthe present invention can also be used as drug for local use.

The pharmaceutical composition of the present invention comprisescantharidic anhydride-essential oils preparation and pharmaceuticallyacceptable vehicle or excipient, wherein the cantharidic anhydride is ina range of 0.05%-0.7% by weight based on total weight of cantharidicanhydride-essential oils preparation.

In addition, the pharmaceutical composition comprising cantharidicanhydride-essential oil has excellent antibacterial and antiviral effecton bacteria such as tubercule bacillus etc and various viruses.

In the pharmaceutical composition of the present invention, cantharidicanhydride is dissolved in essential oils or single constituent ofessential oils to form cantharidic anhydride-essential oil preparation.The cantharidic anhydride has higher solubility in essential oil. Forexample, the solubility of cantharidic anhydride in water isapproximately 0.003%; solubility in fat is 0.05% and solubility inessential oil can reach 0.7%. That is to say, concentration ofcantharidic anhydride in essential oil can be 14 times larger than thatin fat and more than 200 time larger than that in water.

Essential oil is also called as ethereal oil, volatile oil and aromaticoil. It is a fragrant, volatile vegetable oily liquid and is a productobtained by distillation of plant's flower, stem, leave, root or grass.It can also be obtained by the method of pressing, extraction oradsorption. There are many kinds of essential oil. Their mainingredients include terpenes, arenes, alcohols, aldehydes, ketones,ethers, esters, phenols and the like. For example, cinnamon oil contains85%-90% cinnamaldehyde, small amount of cinnamic acid, benzoic acid,coumarin and the like, which have sweet smelling similar to cinnamicaldehyde having slight rosa banksiae fragrance and are used infoodstuff. Citronella oil mainly contains citronellal (35%-50%) andgeraniol (35%-45%), which has smooth and sweet fragrance and slightlemon fragrance and are widely used in perfuming of soaps and articlesfor family uses. Essential oils are soluble in organic solvent such asethanol. Most of them can not be dissolved in water or are slightlysoluble in water.

Therefore, essential oils are used in the present invention forfollowing characteristics: 1. cantharidic anhydride is soluble inessential oil so that the pharmaceutical composition can contain morecantharidic anhydride; 2. Most of essential oils are edible i.e.essential oils used in the present invention are conventionally used andedible, thus it will not have any adverse effect to human body; 3.Essential oils are soluble in alcohols such as ethanol and glycerol.Since ethanol and glycerol are soluble in water, it is possible toprepare water-soluble pharmaceutical composition for local usecomprising cantharidic anhydride-essential oil preparation. Suchpharmaceutical composition has more effectively antiviral andantibacterial effect.

In addition, essential oils with boiling point higher than 100° C.preferably higher than 160° C. are used in the present invention becausethe composition of cantharidic anhydride-essential oil is preferably becarried out under heating condition. Essential oil with higher boilingpoint will not volatilize during the course of preparation. When thepreparation of cantharidic anhydride-essential oil composition iscarried out at relatively lower temperature such as 50° C.-100° C., theessential oil, as far as the boiling point of is higher than 100° C.,can be effectively used. Therefore, most of the conventional edibleessential oils can be used to prepare the composition of cantharidicanhydride-essential oil according to the present invention.

The present invention also provides a method for preparing cantharidicanhydride pharmaceutical composition, comprising

(1) adding cantharidic anhydride in an amount of 0.05%-0.7% by weightinto refined essential oils or single constituent of essential oils,stirring at 40° C.-160° C. under atmospheric pressure and dissolvingcantharidic anhydride in essential oils to obtain cantharidicanhydride-essential oil preparation;

(2) mixing the obtained cantharidic anhydride-essential oil preparationwith pharmaceutically acceptable vehicle or excipient, and stirringhomogeneously to obtain the desired pharmaceutical composition.

The cantharidic anhydride used in the present invention can be preparedaccording to the method described in Chinese Patent 90106365. Inspecifically, the preparation of cantharidic anhydride comprisesfollowing steps:

(1) Soaking cantharis powders in organic solvents to extract cantharidicanhydride into organic phase;

(2) Drying the said cantharis powder having extracted by organicsolvent, Soaking dried powders in solution of concentrated hydrochloricacid and organic solvents, wherein cantharidic acid is converted intocantharidic anhydride by concentrated hydrochloric acid and cantharidicanhydride is dissolved into organic phase by using organic solvents;

(3) Separating cantharidic anhydride and cantharidic oil from organicphase of Step 1, adding petroleum ether to extract cantharidic oil,thereby separating cantharidic anhydride from cantharidic oil, andcombining the cantharidic anhydride obtained with that separated in Step2.

In the above-mentioned method of preparation for cantharidic anhydride,the organic solvent is selected from the group consisted of acetone,dichloromethane, trichloromethane, and ether.

Furthermore, in order to avoid the volatilization of effectiveingredients of the cantharis, the cantharis powders are obtained bypulverizing and drying cantharis at a temperature of 55° C. or less.

According to the conventional method of preparation for thepharmaceutical composition, pharmaceutically acceptable vehicles orexcipients are added to prepare antiviral and antibacterialpharmaceutical compositions of the present invention. Following routinepharmaceutical process, different forms of pharmaceutical compositionscan be prepared by using pharmaceutically acceptable vehicles orexcipients. The pharmaceutically acceptable vehicles or excipients arethose conventionally used in the art and have not special limitation.

The present inventor has already carried out numerous studies regardingthe antiviral and antibacterial effect of the cantharidic anhydride. Thepresent inventor found found that culture medium containing 0.01 ppmcantharidic anhydride can completely deactivate tubercule bacillus andstaphylococcus (the minimum concentration for virus deactivation ofcantharidic anhydride is 0.000132 ppm). It was reported in literaturethat the minimum concentration for tubercule bacillus deactivation ofstreptomycin is 10 ppm, for rifampicin is 25 ppm. The above-mentionedtest indicates that cantharidic anhydride has stronger antibacterialeffect against tubercule bacillus and staphylococcus and thus haspotential value of clinical use.

The cantharidic anhydride-essential oils preparation were used to treatpulmonary tuberculosis and tuberculous pleurisy end achieved much betterclinical effect than those by combining administration of antitubercularagent such as isoniazid, streptomycin and rifampicin. In addition,cantharidic anhydride does not have adverse reaction to liver and bonemarrow as other anti-tubercule bacillus drugs such as rifampicin did.The subjective symptom and blood sedimentation rate is recovered fasterthan those of antitubercular agents.

In addition, chloromycetin eye drops or erythromycin eye ointment areusually used in treating bacterial conjunctivitis. When eye drops orointment comprising cantharidic anhydride (3-6 ppm) essential oilpreparations of the present invention are used, the effective rateapproaches 99%. The symptom mitigates within 1-2 hours afteradministration and heals in about 1-3 days. The cantharidicanhydride-essential oils preparation has much better effects than thoseof antibacterial chloromycetin eye drops or ointment.

When the cream comprising cantharidic anhydride-essential oilpreparations of the present invention are applied to furuncle. The painwill be reduced after 2-3 hours and most of bouton disappear within 2-3days.

Clinical tests have proved that the pharmaceutical compositioncomprising cantharidic anhydride-essential oil preparations has muchimproved clinical effect. Furthermore, it has lower dosage, convenientusage and without any adverse reaction.

In addition, essential oil is used as solvent of cantharidic anhydridein the present invention. Since the amount of essential oil used is nothigh and essential oils without adverse effect are selected, any sideeffects will not occur due to the addition of essential oil. Forpeppermint oil, and peppermint oil, the amount used is determinedaccording to the need of production. In the preparations of the presentinvention, the amounts of sweet orange oil, spearmint oil, carvol,limonene used are far lower than that for food additives as prescribedby Committee for Food Code of United Nations.

Most of the essential oils are easier to be absorbed by skin than fatsdo. Therefore, the use of essential oils as solvents will enhance thepermeability of medicament and thus improve their bioavailability. Themedicament containing essential oil is easier to permeate into outermembrane of bacteria or virus thereby enhances the effect of cantharidicanhydride. Some essential oil will be excreted through lung. Therefore,the effect of cantharidic anhydride as an anti-pulmonary tuberculosismedicament will be promoted. Many kinds of essential oil are easilysoluble in vegetable oil and can prepare extremely effective creams orointments. Some essential oils are easily soluble in glycerol. In thepresent invention, Cantharidic anhydride is first dissolved in essentialoil and then is made into preparations with glycerol. The saidpreparations are suitable for sexual organ use and becomes goodpreparation for controlling infections of hepatitis virus B and C andhuman immunodeficiency virus (HIV).

The present invention will be further described in detail by means offollowing examples. Of course, the present invention is not limited bythese examples.

EXAMPLES Example 1 Preparation of Cantharidic Anhydride-Essential OilPreparation

Cantharidic anhydride-essential oil preparation of the present inventioncan be prepared according to the present inventor's early Chinese Patent(Application Number: 91110970.6). The process of preparation is asfollows:

-   -   (1) Cantharis was pulverized in a pulverizer under 55° C. in dry        state and dried cantharis powder was obtained.    -   (2) Cantharis powder was transferred to a soaking pot and was        soaked with organic solvent such as acetone for 24 hrs or more.        Repeated percolation for several times. The separated organic        phase was charged into separator No. 1 and solid phase was put        into lixiviating pot.    -   (3) Solid phase of cantharis powder residue was transferred to        lixiviating pot was soaked with a solution of concentrated        hydrochloric acid and acetone for 24 hrs or more and repeated        percolation with acetone for many times. Organic phase was        separated and was transferred to separator No. 2 while the        residue was discharged.    -   (4) In separator No. 1, cantharidic anhydride and cantharidic        oil in the organic phase mentioned in step (2) was separated        from organic solvent acetone. Acetone was recovered while        cantharidic anhydride and cantharidic oil was transferred to        separator No. 3    -   (5) In the separator No. 3, petroleum ether was added to        dissolve cantharidic oil. In the separator, cartharidic        anhydride was separated from cantharidic oil. Cantharidic oil        was discharged while cantharidic anhydride was transferred to        refining pot.    -   (6) In the separator No. 2, organic phase was separated from        inorganic phase (concentrated hydrochloric acid not reacted and        salts formed after reaction). After inorganic phase was        discharged, cantharidic anhydride was separated from organic        solvent acetone and was transferred to refining pot.    -   (7) In the refining pot, cantharidic anhydride was washed        repeatedly with petroleum ether, reheated to dissolve with        solvent such as acetone and ethanol, cooled and crystallized to        yield cantharidic anhydride crystal. Finally the said crystals        were repeatedly heated to dissolve in appropriate amount of        acetone. After cooling, crystallization was carried out 2 times        or more, thus pure crystals of cantharidic anhydride with purity        higher than 99% can be obtained. The said pure crystals were        transferred to formulation kettle.    -   (8) Essential oils such as sweet orange oil, peppermint oil,        spearmint oil or single constituent of essential oil such as        limonene, carvol and the like were refined and sterilized. The        refined essential oil was transferred to formulation kettle.    -   (9) In the formulation kettle, cantharidic anhydride and        essential oil charged in a specified proportion were heated and        stirred at 40° C.-180° C. and under atmosphere pressure to        dissolve cantharidic anhydride in essential oil. When        temperature reached about 90° C., cantharidic anhydride was        quickly dissolved in essential oil, if further increasing        temperature, the dissolution rate would not increased as the        temperature. In the course of production, the most preferable        dissolution temperature is in the range of 88-98° C. and the        dissolution process can be completed within 30 minutes. The        cantharidic anhydride-essential oil preparation containing        0.26%-0.7% cantharidic anhydride is obtained according to above        steps 1-9.

To the cantharidic anhydride-essential oil preparation obtained in theabove-mentioned process, a specified proportion of adjuvants (vehicle orexcipient) were added to yield various pharmaceutical compositionscontaining cantharidic anhydride. The manufacturing process was carriedout in formulation kettle. The preparing conditions and the process werefurther described by the following examples.

Example 2 Preparation of Cantharidic Anhydride Cream

The cantharidic anhydride cream was consisted of the followingcomponents:

Cantharidic anhydride 0.15 g-0.25 g Essential oil 40 g-50 g Vegetableoil 15 g-20 g Other matrixes for ointment (stearic acid, 15 g-25 goctadecyl alchol, cetyl alcohol, bee wax, chlorinated vegetable oil andmonostearyl-glyceride) Emulsifiers (tween, sapn, lecithin etc.)   6-20 gbacteriostatic agent appropriate amount water balanced to total 250 g

In the preparation kettle, dispersed emulsion made from distilled waterand other ointment matrixes were melted in cartharidicanhydride-essential oil preparation prepared in example 1. The mixturewas ground in colloid mill to yield emulsion. Upon cooling to roomtemperature, cantharidic anhydride cream was obtained.

Example 3 Preparation of Cantharidic Anhydride Enteric Coating Tablets

The cantharidic anhydride enteric coating tablets:

Cantharidic anhydride 0.06-0.1 g Essential oil  20-30 g Vegetable oil 8-10 g Antioxidant  0.06-0.09 g Other adjuvants appropriate amountTotal amount 300 g (1000 tablets)

In the preparation kettle, a specified amount of adjuvants was added andstirred homogeneously to prepare the tablets only adjuvants, then thecantharidic anhydride-essential preparation obtained in example 1 washomogeneously permeated into the tablets at 40° C.-60° C. Enteric coatedto obtain the cantharidic anhydride enteric coating tablets.

Example 4 Preparation of Intravenous Injection Emulsion of CantharidicAnhydride

Pure cantharidic anhydride 0.08-0.1 g   Refined single constituent ofessential oil 16-20 g  (such as limonene) Refined vegetable oil 80-100 gEmulsifier (lecithin for injection use etc.) 6-12 g Propanetriol 0-20 gWater (for injection use) balanced to 1000 ml

In the preparation kettle, a specified amount of purified water,emulsifier, glycerol were added under nitrogen atmosphere to makeemulsion containing emulsifier. The cantharidic anhydride-essential oilpreparation obtained in example 1 was added and was triturated bycolloid mill and homogenizer to yield highly homogeneous emulsion. Aftersterilization, injection emulsion was obtained. The preparation processfor injection emulsion is carried out under nitrogen atmosphere.

Example 5 Preparation of Cantharidic Anhydride Drop Pills

The cantharidic anhydride drop pill:

Cantharidic anhydride 0.06-0.1 g  Essential oil 22-32 g Vegetable oil10-12 g Antioxidant 0.06-0.09 g Other adjuvants appropriate amount Totalamount 300 g (1000 drop pills)

To the preparation kettle, vegetable oil and other adjuvants were addedand then cantharidic anhydride-essential oil preparation prepared inexample 1 was added and stirred at 66° C.-76° C. for 30-60 minutes toyield oily solution. Dropped to prepare pill, dried, and packed.

Beside Examples 1-5, the cantharidic anhydride-essential oil preparationobtained in Example 1 can also be used to make cantharidic anhydrideinjection, film coating, liniment, patch, rubber plaster, oral emulsion,capsule, enteric coating capsule, soft capsule, drop pill, honey bolusesand the like.

In addition, the cantharidic anhydride-essential oil preparationobtained in Example 1 can also be used to local administration, forexample in the form of sterilization and hygiene agent for treating orpreventing bacterial or viral diseases.

For instance, the following preparations can be prepared:

1. Cantharidic Anhydride Ointment

Cantharidic anhydride 0.0003-0.03 g  Essential oil  6-12 g Fat  460-520g Other ointment matrixes appropriate amount (such as stearic acid,stearyl alcohol, bee wax) Antioxidant 0.2-8 g Total 600 g

To the preparation kettle, sterilized fat and other auxiliary matrixwere added and then cantharidic anhydride-essential oil preparationobtained in example 1 was added. Stirred continuously 1-2 hr at 66-76°C. until completely homogeneity. Filtered, cooled to room temperature,packed to obtain Cantharidic anhydride ointment. The ointment can beused to inhibit completely infections from burns, trauma and stitchedwounds from surgical operations. It has excellent effect of easing pain.The pain can be alleviated even in the case of serious burn.

2. Cantharidic Anhydride Eye Ointment

Cantharidic anhydride 0.0002 g-0.02 g  Essential oil  2-8 g Fat 460g-520 g Other matrixes (such as lanolin, bee wax, appropriate amountliquid paraffin) Antioxidant 0.1 g-8 g  Final total amount 600 g

To the preparation kettle, cantharidic anhydride-essential oil obtainedin Example 1 was added and the sterilized fat and other matrixes wereadded. The eye ointment was prepared by using the same method forpreparing ointment. The said eye ointment had excellent effect fortreating bacterial and viral infection of conjunctiva, which was betterthan that of the prior antibacterial and antiviral eye ointments.

3. Cantharidic Anhydride Emulsion for Special Use

Components:

Cantharidic anhydride 0.0003-0.03 g  Essential oil 3-9 g Antioxidant0.1-0.9 g Glycerol (can replaced by propylene glycol, appropriate amountsorbitol, polyethylene glycol) Other adjuvants and perfume appropriateamount Distilled water appropriate amount Total 900 g

Glycerol and other adjuvants were added and then cantharidicanhydride-essential oil preparation obtained in Example 1 was added intothe preparation kettle No. 10. Stirred continuously for 30-60 minutes at66-76° C. and then distilled water and perfume were added and stirredagain at 66-76° C. for 10-20 minutes. After cooled to room temperature,the desired slightly viscous emulsion for special use was obtained. Theemulsion could be used for sexual organ to deactivate completely humanimmunodeficiency virus, hepatitis B virus, hepatitis C virus and variousbacteria. It can prevent sexual infections. It also make patient feelcomfortable and refreshing. Since the concentration of cantharidicanhydride exceeds the minimum deactivation concentration of virus over10,000 times and cantharidic anhydride has no side effect to humanorgans, the emulsion is an ideal medicament for preventing sexualinfectious diseases.

4. Cantharidic Anhydride Sterilized Wipes

Cantharidic anhydride 0.00036-0.036 g  Essential oil (such as phenethylalcohol) 3.6-9.6 g Antioxidant  2-12 g Glycerol appropriate amount Otheradjuvants and perfume appropriate amount Distilled water,skin-protecting solution appropriate amount of Chinese traditionalmedicine) total 900 g

To the preparation kettle, glycerol and other adjuvants were added andthen cantharidic anhydride-essential oil preparation obtained in Example1 was added. Stirred continuously for 30-60 minutes at 66-76° C., thendistilled water and perfume were added and stirred again at 66-76° C.for 10-20 minutes. The drug solution for skin-protecting anddisinfectant was obtained.

Wipes were soaked in the said drug solution according to specifiedweight proportion and the soaked drug solution was packed and sealed toobtain the final product.

The drug concentration of the wipes after the vaporization of watercould reach over 30,000 times of the minimum viral deactivationconcentration of cantharidic anhydride. The wipes can kill virus andbacteria on skin and on the superficial opening of wound of skin.Meantime, the wipes have the effect of wetting, protecting, and cleaningskin. In a trip or outdoors, it could be used to wash or clean face andhand. In case of trauma, it could be used to urgently wrap or cover thewound. It is an ideal article of sanitation and health care during trip.

Using the method similar to that above-mentioned, various local beprepared such as rubber plaster, aerosol, eye dropping oils, oils forburn, spray for burn, sterilization spray, suppositories, eye drops, eardrops, nose drops, sterilizing solution for oral cavity and the like.

Example 6 Clinical Experiments Concerning Tubercule Bacillus andHepatitis Virus of the Pharmaceutical Composition Comprising CantharidicAnhydride-Essential Oil Preparation

The course of chemotherapeutic treatment for tuberculosis by combinationof isoniazid etc has been prolonged to 18-24 months for “standardchemotherapeutic treatment” and 6-9 months for “short chemotherapeutictreatment” by the Chinese health administration. This suggests that thecuring effect of the drug for treating tuberculosis has lowered. As aresult, long time treatment is needed. The percentage of occurrence ofside effect after above-mentioned long time combining administration hasreached 70%-80%. In the period of administering drug of 1-2 year,patients were unable to take part in family and social labors mainly dueto side effects of drug. However large amount of clinical experimentsusing pharmaceutical compositions comprising cantharidicanhydride-essential oil preparation as the main ingredient according tothe present invention have ideal curative effects. For instance, whencomposition according to Example 2 was used to treat tuberculouspleuritis, time of auto-absorption rate of accumulated chest fluid, bodytemperature, blood sedimentation rate were all significantly better thanthat achieved by combination of isoniazid, streptomycin and rifampicin.In addition, the cantharidic anhydride preparations did not have anyside effects. The specific clinical experiments were carried outaccording to the following processes.

-   -   1. The patients suffering with pulmonary tuberculosis in control        group (I) were subjected to conventional combined treatment of        isoniazid, streptomycin and rifampicin. The patients took drug        continuously for 180 days and the results of clinical treatment        were observed.    -   2. The patients suffering with pulmonary tuberculosis in control        group (II) were administrated fat soluble cantharidic anhydride        emulsion (preparation process was similar to Example 2 except        essential oil was replaced by fat, the dosage of cantharidic        anhydride in the pharmaceutical composition is identical with        that of the following treatment group). The patients took drug        continuously for 90 days and the results of clinical treatment        were observed.    -   3. The patients suffering with pulmonary tuberculosis in treated        group were administrated emulsion prepared in Example 2. The        patients took drug continuously for 90 days and the results of        clinical treatment were observed.

In addition, for control group (II) and treatment group, the emulsionswere applied on the chest and back of the patients 3 times a day and 2 gof emulsion each time. The results obtained for the above treatmentswere illustrated in Tables 1-3.

TABLE 1 Comparison of three treatments for patients suffering frominfiltrative pulmonary tuberculosis Item The symptom such as The bodyanepithymia temperature Getting and and blood better recoveryhypodynamia sedimentation shown by shown by disappearing rate X-rayX-ray No. of (Average recovery diagnosis diagnosis Group cases days)(Av. Days) (Av. Day) (Av. Days) Remarks Control 36 Over 60% 72.6 54.292.6 days Take drug group (I) patients did 26 cases continuously notrecover recovered, for 180 days due to side 10 cases effects can notrecovered Control 22 16.2 26.3 48.2 62.3 Take drug group (II)continuously for 90 days Treatment 26 11.6 19.8 19.8 43.3 Take druggroup continuously for 90 days

TABLE 2 Comparison of three treatments for patients suffering fromtuberculous pleurisy Item The symptom The body such as temperatureAccumlated anepithymia and chest Accumulated and blood fluid chest fluidhypodynamia sedimentation decreased completely disappearing rate shownby absorbed No. of (Average recovery CT shown by CT Group cases days)(Av. Days) (Av. days) (Av. days) Remarks Control 30 Over 60% 59.6 42.366.6 Take drug group (I) patients did continuously not recover for 90days due to side effects Control 20 13.2 24.6 24.6 38.2 Take drug group(II) continuously for 90 days Treatment 22  9.9 14.6 14.6 31.6 Take druggroup continuously for 90 days

TABLE 3 Comparison of occurrence of side effects for three treatmentsfor patients suffering with infiltrative pulmonary tuberculosis andtuberculous pleurisy(3 months) side effect The symptom Increase ofDecrease such as liver toxic of anepithymia, glutamate marrow eatingless pyruvate and and nausea transaminase leucocyte Other side No. ofGroup (%) (%) (%) effects cases Control 82.6 36.2 69.3 0 66 group (I)Control 0 0 0 0 42 group (II) Treatment 0 0 0 0 48 group

In addition, clinical experiments were carried out on patients sufferinghepatitis, especially those infected with hepatitis AE virus andhepatitis BC virus by using the pharmaceutical composition of thepresent invention. Three groups were carried out at same time.

-   -   1. In control group (I), decoction of traditional Chinese        medicine was used,    -   2. In control group (II), fat-soluble cantharidic anhydride        emulsion was used,    -   3. In treatment group, the emulsion comprising cantharidic        anhydride prepared in Example 2 was used.

In the above control group (II) and treatment group, the same dosage andadministrating methods were used. The results of three groups wereillustrated in Table 4.

TABLE 4 Comparison of the curative effects on patients infected withhepatitis AE virus and hepatitis BC virus. Item Jaundice Symtoms ofRecovered anepithymia and Symptoms shown by hypodynamia completely GPT,ALT getting better disappear analysis group (Av. Days) (Av. Days) (Av.Days) No. of cases Control Hepatitis AE 6.8 31.8 36.3 16 group (I) virusgroup Hepatitis BC 21.2 38.6 56.6 20 virus group Control Hepatitis AE3.6 12.6 21.1 20 group (II) virus group Hepatitis BC 11.2 22.3 32.3 22virus group Treatment Hepatitis AE 2.6 10.8 16.2 26 group virus groupHepatitis BC 8.3 19.6 28.2 26 virus group

In addition, results of clinical experiments obtained in treatments ofacute viral conjunctivitis by using eye dropping oils of fat solublecantharidic anhydride and cantharidic anhydride-essential oil of thepresent invention was illustrated in Table 5.

TABLE 5 Comparison of curative effects of eye dropping oils of fatsoluble cantharidic anhydride and cantharidic anhydride-essential oil onacute viral conjunctivitis Item Symptom of avoiding Symptom of light,shedding tears reddish swelling and sand-rubbing disappeared Grouplighten completely P value Treatment group Av. 22 hrs Av. 46.8 hrs usingeye oil of fat soluble cantharidic anhydride Treatment group Av. 32.2minutes Av. 26 hrs < 0.01 using eye oil of cantharidicarihydride-essential oil

Example 7 Antiviral Experiment Based on Tissue Culture Technique ofCantharidic Anhydride-Essential Oil Preparation

Cantharidic anhydride antiviral test solution:

Cantharidic anhydride 0.1 g Essential oil (completely soluble inglycerol)  30 g Glycerol appropriate amount Distilled water appropriateamount Total 1000 g  

The contration of cantharidic anhydride of Cantharidic anhydrideantiviral test solution was 100 ppm. According to the scheme of tissueculture, it is demanded that each group is cultured in different drugconcentration and then diluted with water. The Cantharidic anhydrideantiviral test solution was used as treatment group and cantharidicanhydride directly dissolved and diluted by water was used as controlgroup. The experimental results are demonstrated in Table 6.

TABLE 6 Comparison of the antiviral experiment results on eight kinds ofvirus by cantharidic anhydride-essential oils preparation and aqueoussolution of cantharidic anhydride Minimum virus Minimum virusdeactivation deactivation concentration concentration of of cantharidiccantharidic anhydride anhydride for for treatment Kind of virus controlgroup (ppm) group (ppm) Adenovirus III 0.000163 Cyncytial virus 0.000489Coxsackie B5 virus 0.000132 Pollomyelitis virus 0.000256 Herpes virus0.000322 Rotavirus 0.000169 Hepatitis B virus 0.0028 Human 0.0072immunodeficiency virus

In summary, the pharmaceutical compositions comprising cantharidicanhydride-essential oils preparation according to the present inventionhave broad spectrum of antiviral and antibacterial as shown by theresult of a number of clinical experimental results. The pharmaceuticalcompositions according to the present invention have excellent curativeeffect on tuberculous disease, viral hepatitis, viral enteritis and thelike.

1. An antiviral and antibacterial pharmaceutical composition comprisingcantharidic anhydride as the effective ingredient and a pharmaceuticallyacceptable vehicle or excipient, wherein the cantharidic anhydride isdissolved in essential oils or a single constituent of essential oils at40-160° C. to form a cantharidic anhydride-essential oils preparation,wherein the essential oils are selected from the group consisting ofethereal oil, volatile oil and aromatic oil, and the cantharidicanhydride is dissolved in a range of 0.05%-0.7% by weight based on atotal weight of the cantharidic anhydride-essential oils preparation. 2.An antiviral and antibacterial pharmaceutical composition for externallocal use comprising cantharidic anhydride as the effective ingredientand a pharmaceutically acceptable vehicle or excipient, wherein thecantharidic anhydride is dissolved in essential oils or a singleconstituent of essential oils at 40-160° C. to form a cantharidicanhydride-essential oils preparation, wherein the essential oils areselected from the group consisting of ethereal oil, volatile oil andaromatic oil, and the cantharidic anhydride is dissolved in a range of0.05%-0.7% by weight based on a total weight of the cantharidicanhydride-essential oils preparation.
 3. A pharmaceutical compositionagainst tubercule bacillus comprising cantharidic anhydride as theeffective ingredient and a pharmaceutically acceptable vehicle orexcipient, wherein the cantharidic anhydride is dissolved in essentialoils or a single constituent of essential oils at 40-160° C. to form acantharidic anhydride-essential oils preparation, wherein the essentialoils are selected from the group consisting of ethereal oil, volatileoil and aromatic oil, and the cantharidic anhydride is dissolved in arange of 0.05%-0.7% by weight based on a total weight of the cantharidicanhydride-essential oils preparation.
 4. A pharmaceutical compositionagainst hepatitis virus and enteritis virus comprising cantharidicanhydride as the effective ingredient and a pharmaceutically acceptablevehicle or excipient, wherein the cantharidic anhydride is dissolved inessential oils or a single constituent of essential oils at 40-160° C.to form a cantharidic anhydride-essential oils preparation, wherein theessential oils are selected from the group consisting of ethereal oil,volatile oil and aromatic oil, and the cantharidic anhydride isdissolved in a range of 0.05%-0.7% by weight based on a total weight ofthe cantharidic anhydride-essential oils preparation.
 5. Thepharmaceutical composition according to claim 1, wherein the selectedessential oils are edible essential oils.
 6. A method for preparing thecantharidic anhydride pharmaceutical composition according to claim 1,comprising: (1) adding cantharidic anhydride in an amount of 0.05%-0.7%by weight into the essential oils or a single constituent of theessential oils, stiffing at 40-160° C. under atmospheric pressure, anddissolving the cantharidic anhydride in the essential oils to obtain thecantharidic anhydride-essential oils preparation; (2) mixing theobtained cantharidic anhydride-essential oils preparation with thepharmaceutically acceptable vehicle or excipient, and stiffinghomogeneously to obtain the desired pharmaceutical composition.
 7. Themethod according to claim 6, wherein the cantharidic anhydride isprepared according to the following steps: (1) Soaking cantharis powdersin organic solvents to extract cantharidic anhydride into an organicphase; (2) Drying the cantharis powder extracted by organic solvent,soaking the dried powders in a solution of concentrated hydrochloricacid and organic solvents, wherein cantharidic acid is converted intocantharidic anhydride by the concentrated hydrochloric acid, andcantharidic anhydride is dissolved into the organic phase by using theorganic solvents; (3) Separating the cantharidic anhydride andcantharidic oil from the organic phase of Step 1, adding petroleum etherto extract the cantharidic oil, thereby separating the cantharidicanhydride from the cantharidic oil, and combining the cantharidicanhydride obtained with that separated in Step
 2. 8. The methodaccording to claim 7, wherein the organic solvent is selected from thegroup consisted of acetone, dichloromethane, trichloromethane, andether.
 9. The method according to claim 7, wherein the cantharis powdersare obtained by pulverizing and drying cantharis at a temperature of 55°C. or less.
 10. The pharmaceutical composition according to claim 2,wherein the selected essential oils are edible essential oils.
 11. Amethod for preparing the cantharidic anhydride pharmaceuticalcomposition according to claim 2, comprising: (1) adding cantharidicanhydride in an amount of 0.05%-0.7% by weight into the essential oilsor a single constituent of the essential oils, stiffing at 40-160° C.under atmospheric pressure, and dissolving cantharidic anhydride in theessential oils to obtain the cantharidic anhydride-essential oilpreparation; (2) mixing the obtained cantharidic anhydride-essential oilpreparation with the pharmaceutically acceptable vehicle or excipient,and stiffing homogeneously to obtain the desired pharmaceuticalcomposition.
 12. The method according to claim 11, wherein the organicsolvent is selected from the group consisted of acetone,dichloromethane, trichloromethane, and ether.
 13. The method accordingto claim 11, wherein the cantharis powders are obtained by pulverizingand drying cantharis at a temperature of 55° C. or less.
 14. Thepharmaceutical composition according to claim 3, wherein the selectedessential oils are edible essential oils.
 15. A method for preparing thecantharidic anhydride pharmaceutical composition according to claim 3,comprising: (1) adding cantharidic anhydride in an amount of 0.05%-0.7%by weight into the essential oils or a single constituent of theessential oils, stiffing at 40-160° C. under atmospheric pressure, anddissolving cantharidic anhydride in the essential oils to obtain thecantharidic anhydride-essential oil preparation; (2) mixing the obtainedcantharidic anhydride-essential oil preparation with thepharmaceutically acceptable vehicle or excipient, and stiffinghomogeneously to obtain the desired pharmaceutical composition.
 16. Themethod according to claim 15, wherein the organic solvent is selectedfrom the group consisted of acetone, dichloromethane, trichloromethane,and ether.
 17. The method according to claim 15, wherein the cantharispowders are obtained by pulverizing and drying cantharis at atemperature of 55° C. or less.
 18. The pharmaceutical compositionaccording to claim 4, wherein the selected essential oils are edibleessential oils.
 19. A method for preparing the cantharidic anhydridepharmaceutical composition according to claim 4, comprising: (1) addingcantharidic anhydride in an amount of 0.05%-0.7% by weight into theessential oils or a single constituent of the essential oils, stiffingat 40-160° C. under atmospheric pressure, and dissolving cantharidicanhydride in the essential oils to obtain the cantharidicanhydride-essential oil preparation; (2) mixing the obtained cantharidicanhydride-essential oil preparation with the pharmaceutically acceptablevehicle or excipient, and stiffing homogeneously to obtain the desiredpharmaceutical composition.
 20. The method according to claim 19,wherein the organic solvent is selected from the group consisted ofacetone, dichloromethane, trichloromethane, and ether.
 21. The methodaccording to claim 19, wherein the cantharis powders are obtained bypulverizing and drying cantharis at a temperature of 55° C. or less.